Method for the production of 2,4-dihydroxybutyric acid

ABSTRACT

Methods for the production of 2,4-dihydroxybutyrate (2,4-DHB) from erythrulose and other four-carbon sugars are disclosed. The improved methods facilitate the production of 2,4-DHB that is a precursor for biorenewable and animal nutrition chemicals among others.

This application is a continuation application of U.S. application Ser.No. 15/738,249, filed Dec. 20, 2017, which is a 371 national phase ofPCT/US2016/039282, filed Jun. 24, 2016, and claims benefit under 35U.S.C. § 119(e) of U.S. Provisional Application No. 62/204,796, filedAug. 13, 2015, and of U.S. Provisional Application No. 62/184,571, filedJun. 25, 2015, the disclosures of which are incorporated, in theirentirety, by this reference. The invention relates to the production of2,4-dihydroxybutyrate from erythrose and other four-carboncarbohydrates.

TECHNICAL FIELD Background

2,4-dihydroxybutyrate (also referred to as 2,4-DHB or DHB) is a highlyuseful chiral intermediate and is of considerable economic interest. DHBcan be readily converted into α-hydroxy-γ-butyrolactone in aqueous mediaby adjusting to the appropriate pH. α-hydroxy-γ-butyrolactone is aprominent precursor for the production of the methionine substitute2-hydroxy-4-(methylthio)-butyrate (HMTB), as described in U.S. PatentApplication 2009/0318715, which has a large market in animal nutrition.DHB is also a promising precursor for biorenewable chemicals such as3-hydroxypropanal, 3-hydroxypropionic acid, 3-propanediol, and malonicacid. DHB has been produced using complex metabolic engineeringapproaches as described in U.S. Patent Application Publication No.2013/0273623, European Patent Application Publication Nos. 2841584 A2and 2872640 A1. Such metabolic engineering approaches require expensiveraw materials and complex reaction conditions. Other synthetic routeshave used expensive raw materials such HMTB as described in U.S. PatentApplication Publication No. 2013/0204016 A1. There remains a need forcost-effective methods to produce DHB.

The reaction of sugars in alkaline conditions have been studied sincethe nineteenth century. Sugars react with hydroxide in complicatedpathways both in the presence of oxygen and in anaerobic conditions. Forexample, glucose or fructose react with oxygen gas in an alkaline watersolution (see for example, as described in U.S. Pat. Nos. 4,125,559 and5,831,078), where the 1-2 carbon bond is broken, yielding predominantlyformic acid (from carbon 1) and arabinonic acid (from carbons 2-6).Also, large amounts of shorter carbon chain acids also are produced asdescribed in Tapani Vuorinen, “Cleavage of the IntermediateHydroperoxides in the Oxidation of D-Glucose and D-Fructose withOxygen,” Carbohydrate Research, 141 (1985): 319-332. Anaerobic reactionsof sugars in alkaline conditions are generally termed degradations andresult in complex mixtures of reaction products that are difficult toanalyze but that include small amounts of DHB, as described in Byung YunYang and Rex Montgomery, “Alkaline Degradation of Glucose: Effect ofInitial Concentration of Reactants,” Carbohydrate Research 280 (1996):27-45, and J. F. Harris, “Alkaline Decomposition of D-Xylose-1-14C,D-Glucose-1-14C, and D-Glucose-6-14C,” Carbohydrate Research 23 (July1972): 207-215. Moreover, compounds such as Class I Caramel Color arecreated by reacting glucose with hydroxide in anaerobic conditions.Under milder alkaline conditions glucose is known to merely isomerizeinto fructose, as described in U.S. Pat. No. 3,256,270. Because oxygenis sparingly soluble in water, often degradations occur when open to theatmosphere.

There remains a need in the art for cost effective methods for theproduction of DHB from four carbon sugars such as erythrose. Erythroseitself is a rare four carbon sugar that has recently been produced on alarge scale via electrochemical decarboxylation as described in U.S.Patent Application Publication No. 2007/0181437. The present disclosureprovides a method of converting four-carbon sugars, to four-carbon DHBby reacting the sugars in an alkaline solution.

SUMMARY

In one aspect, a method of producing 2,4-dihydroxybutyrate is disclosedwhich includes mixing a four carbon sugar and a hydroxide salt insolution. In some embodiments, the four carbon sugar may be threose orerythrulose. In some embodiments, the temperature of the solution ismaintained below 100° C. In some embodiments, the four carbon sugar isdiluted sufficiently to result in a molar yield of DHB that is greaterthan 40%. The four carbon sugar may be diluted with a solutioncontaining DHB. In some embodiments, the method is carried out in acontinuous reactor system. In some embodiments, the hydroxideconcentration of the solution may be between about 0.1 M and about 4 M.

In some embodiments, the method includes removing oxygen from thesolution. The oxygen may be removed by venting the solution with a gasselected from nitrogen, argon, and mixtures of the same. The oxygen mayalso be removed by venting the solution with hydrogen.

In some embodiments, the four carbon sugar is erythrose. The erythrosemay be diluted with a solution containing one or more other organic acidsalts. The erythrose may be diluted with a solution containing DHB.

DETAILED DESCRIPTION Definitions

“Erythrose” refers to an aldose (tetrose) carbohydrate aldehyde withchemical formula C₄H₈O₄, including any stereoisomers, derivatives andanalogs thereof. Unless otherwise indicated, recitation of “erythrose”herein is intended to include, without limitation, the molecules:D-(−)-erythrose, L(+)-erythrose, D-erythrose, L-erythrose, andmeso-erythrose. A Fischer Projection of the D-erythrose structure (1) isshown below.

“Threose” refers to an aldose (tetrose) carbohydrate aldehyde withchemical formula C₄H₈O₄, including any stereoisomers, derivatives andanalogs thereof. Unless otherwise indicated, recitation of “threose”herein is intended to include, without limitation, the molecules:D-(−)-threose, L(+)-threose, D-threose, L-threose, and meso-threose.

“Erythrulose” refers to a ketose (tetrulose) carbohydrate ketone withchemical formula C₄H₈O₄, including any stereoisomers, derivatives andanalogs thereof. Unless otherwise indicated, recitation of “erythrulose”herein is intended to include, without limitation, the molecules:D-(−)-erythrulose, L(+)-erythrulose, D-erythrulose, L-erythrulose.

“2,4-dihydroxybutyrate” (also known as 2,4-DHB or DHB) is an organicacid and refers to the carbohydrate tetronic acid with the chemicalformula C₄H₈O₄, or salt thereof, including any stereoisomers,derivatives and analogs thereof. Unless otherwise indicated, recitationof “2,4-dihydroxybutyrate” (also known as 2,4-DHB or DHB) herein isintended to include, without limitation, the molecules:(S)-2,4-dihydroxybutyrate, (R)-2,4-dihydroxybutyrate, meso2,4-dihydroxybutyrate, and 3-deoxytetronate.

As used herein, “derivative” refers to a chemically or biologicallymodified version of a chemical compound that is structurally similar toa parent compound and (actually or theoretically) derivable from thatparent compound. A derivative may, or may not, have different chemicalor physical properties of the parent compound. For example, thederivative may be more hydrophilic or it may have altered reactivity ascompared to the parent compound. Derivatization (i.e., modification) mayinvolve substitution of one or more moieties within the molecule (e.g.,a change in functional group) that do not substantially alter thefunction of the molecule for a desired purpose. The term “derivative” isalso used to describe all solvates, for example hydrates or adducts(e.g., adducts with alcohols), active metabolites, and salts of theparent compound. The type of salt that may be prepared depends on thenature of the moieties within the compound. For example, acidic groups,for example carboxylic acid groups, can form, for example, alkali metalsalts or alkaline earth metal salts (e.g., sodium salts, potassiumsalts, magnesium salts and calcium salts, and also salts quaternaryammonium ions and acid addition salts with ammonia and physiologicallytolerable organic amines such as, for example, triethylamine,ethanolamine or tris-(2-hydroxyethyl)amine). Basic groups can form acidaddition salts, for example with inorganic acids such as hydrochloricacid, sulfuric acid or phosphoric acid, or with organic carboxylic acidsand sulfonic acids such as acetic acid, citric acid, benzoic acid,maleic acid, fumaric acid, tartaric acid, methanesulfonic acid orp-toluenesulfonic acid. Compounds which simultaneously contain a basicgroup and an acidic group, for example a carboxyl group in addition tobasic nitrogen atoms, can be present as zwitterions. Salts can beobtained by customary methods known to those skilled in the art, forexample by combining a compound with an inorganic or organic acid orbase in a solvent or diluent, or from other salts by cation exchange oranion exchange.

As used herein, “analogue” refers to a chemical compound that isstructurally similar to another but differs slightly in composition (asin the replacement of one atom by an atom of a different element or inthe presence of a particular functional group), but may or may not bederivable from the parent compound. A “derivative” differs from an“analogue” in that a parent compound may be the starting material togenerate a “derivative,” whereas the parent compound may not necessarilybe used as the starting material to generate an “analogue.”

Any concentration ranges, percentage range, or ratio range recitedherein are to be understood to include concentrations, percentages orratios of any integer within that range and fractions thereof, such asone tenth and one hundredth of an integer, unless otherwise indicated.Also, any number range recited herein relating to any physical feature,such as polymer subunits, size or thickness, are to be understood toinclude any integer within the recited range, unless otherwiseindicated. It should be understood that the terms “a” and “an” as usedabove and elsewhere herein refer to “one or more” of the enumeratedcomponents. For example, “a” polymer refers to one polymer or a mixturecomprising two or more polymers. As used herein, the term “about” refersto differences that are insubstantial for the relevant purpose orfunction.

Alkaline Conversion

The process of converting four carbon sugars to DHB is described below.In some embodiments, a four carbon sugar can be provided as a solutionto which hydroxide ion is added in the form of an alkali metal, alkalineearth metal, or ammonium salt, or salt solution. The yield of DHB inthese reactions is greatly influenced by the concentration of the sugarand the concentration of hydroxide. At a given temperature, reduced fourcarbon sugar concentration results in increased DHB yields, andincreased hydroxide concentration results in increased DHB yields. Atthe same hydroxide concentration, increasing the temperature results inincreased DHB yields. In some embodiments, erythrose can be provided asthe four carbon sugar. In other embodiments, threose or erythrulose canbe provided as the four carbon sugar. In one embodiment, the solution ispurged with a gas such as nitrogen, hydrogen, or argon or mixtures ofthe same to remove oxygen from the solution.

In some embodiments, the alkaline conversion of a four carbon sugar intoDHB by introducing a solution of the four carbon sugar into a continuousreactor holding sufficient solution to dilute the four carbon sugar toresult in high yields of DHB. In such a reactor, the four carbon sugarwould be diluted by a solution contain hydroxide salt, DHB, and/or otherorganic acid salts. The reactor includes the means to maintain thesolution at specific temperature, and the means to introduce thesolution of the four carbon sugar, a solution of hydroxide salt, and themeans to remove product.

EXAMPLES Example 1

A 153 gram erythrose per liter solution in water was provided. Table 1provides the results of experiments where the indicated volume of theerythrose solution was add to 10 ml of 1 M sodium hydroxide in water.The mixtures were stirred for 60 min at the indicated temperature.

TABLE 1 Erythrose Yield (DHB) Sample Solution Temp on erythrose ID Vol.(mL) (C.) % 921-91-3 1 22 16% 921-91-4 2 22 12% 921-91-5 5 22  2%921-93-1 1 40 57% 921-93-2 2 40 34% 921-93-3 5 40 13% 921-95-1 1 50 64%921-95-2 2 50 44% 921-95-3 5 50 18% 921-95-4 1 60 56% 921-95-5 2 60 41%921-95-6 5 60 20%

Example 2

A 156 gram erythrose per liter solution in water was provided. 1 mL ofthe erythrose solution was added to 100 mL of the indicated hydroxidesolution in Table 2 and stirred at 40° C. for 60 minutes.

TABLE 2 2,4-DHB Exp. Caustic Yield 953-31-4 4M KOH 59% 953-33-1 4M NaOH58% 953-33-3 3.5M LiOH 53%

Example 3

A 153 gram erythrose per liter solution in water was provided. 1 mL ofthe erythrose solution was add to 100 mL of a 4 M sodium hydroxidesolution that also had a concentration of the organic acid salt sodiumarabonate of 2.4 M. The solution mixed for 60 minutes at 40 C. DHB wasquantified showing a molecular yield of greater than 58%.

Example 4

A 100 ml reactor was provided containing 4 M NaOH and heated to 50° C. A80 gram erythrose per liter solution in water was provided. 1 mL of theerythrose solution was added to the reactor per minute, and 45% NaOH wasadded at a rate of 0.19 mL per minute. 1 ml of the erythrose solutionwas added to a 4 M sodium hydroxide solution and stirred under argon.The solution was maintained at 30° C. for 60 min. The reactor volume wasmaintained constant throughout the experiment. After 4 hours thesolution in the reactor was analyzed by HPLC and DHB was quantifiedshowing a molecular yield of 46%.

Example 5

1 L high pressure reactor with headspace entraining hollow-shaft mixerwas provided. 700 mL of a 4 M NaOH solution was provided and the reactorwas heated to 50° C. The reactor was then pressurized with the gasesoutlined in Table 3 to 750 psi. 7 mL of a 136 g per L erythrose solutionwas then added to the reactor. After 60 minutes, the solutions were thenanalyzed by HPLC.

TABLE 3 Gas DHB % Yield Air 34% Oxygen  0% Hydrogen 50%

Example 6

100 mg of threose was added to 50 mL of a 4 M sodium hydroxide solutionand stirred at 40° C. for 60 min. DHB was quantified showing a molecularyield of greater than 56%.

Example 7

A 152 gram erythrulose per liter solution in water was provided. 1 mL ofthe erythrulose solution was added 100 mL of 2M NaOH solution. Thesolution was then stirred at 50° C. for 15 min. DHB was quantifiedshowing a molecular yield of 65%.

Example 8

A 136 gram erythrose per liter solution in water was provided. 1 mL ofthe erythrose solution was added 200 mL of 4 M NaOH solution under ahydrogen headspace. The solution was then stirred at 40° C. for thetimes indicated in Table 4.

DHB yields are also reported in Table 4.

TABLE 4 DHB % Sample Min yield 953-66-2   2 12% 953-66-3   5 30%953-66-4  10 47% 953-66-6  35 58% 953-66-7  60 60% 953-66-8 104 64%

Example 9

A 132 gram erythrose per liter solution in water was provided. 1 mL ofthe erythrose solution was added 100 mL of 1 M BaOH. The solution wasthen stirred at 60° C. for 100 min. DHB was quantified showing amolecular yield of 20%.

Example 10

A 135 gram erythrose per liter solution in water was provided. 1 mL ofthe erythrose solution was added 100 mL of NaOH solution with aconcentration indicated in Table 5. The solution was then stirred at 50°C. for 35 min. DHB was quantified showing a molecular yield indicated inTable 5.

TABLE 5 M Yield Sample NaOH DHB 953-70-4 1 41% 953-70-13 2 54%

Example 11

A 135 gram erythrose per liter solution in water was provided. 1 mL ofthe erythrose solution was added to 100 mL of saturated Lead Hydroxidesolution. The solution was then stirred at 50° C. for 60 min. DHB wasquantified and no DHB was present in the reaction solution.

It will be evident to those skilled in the art that the invention is notlimited to the details of the foregoing illustrative examples and thatthe present invention may be embodied in other specific forms withoutdeparting from the essential attributes thereof, and it is thereforedesired that the present embodiments and examples be considered in allrespects as illustrative and not restrictive, reference being made tothe appended claims, rather than to the foregoing description, and allchanges which come within the meaning and range of equivalency of theclaims are therefore intended to be embraced therein.

STATEMENTS

1. A method of producing 2,4-dihydroxybutyrate, comprising: mixing afour carbon sugar and an hydroxide salt in solution.

2. The method of 1, wherein the four carbon sugar is erythrose.

3. The method of 1, wherein the four carbon sugar is threose orerythrulose.

4. The method of any one of 1-3, wherein the hydroxide concentration ofthe solution is between 0.1 M and 4 M.

5. The method of any one of 1-4, wherein the temperature of the solutionis maintained below 100° C.

6. The method of any one of 1-5, wherein the four carbon sugar isdiluted sufficiently to result in a molar yield of DHB that is greaterthan 40%.

7. The method of any one of 1-6, wherein the four carbon sugar isdiluted with a solution containing DHB.

8. The method of any one of 1-7, wherein the erythrose is diluted with asolution containing one or more other organic acid salts.

9. The method of any one of 7-8, wherein the method is performed in acontinuous reactor system.

10. The method of any one of 1-9, further comprising removing oxygenfrom the solution.

11. The method of 10, wherein oxygen is removed by venting the solutionwith a gas selected from the group consisting of: nitrogen, argon, andmixtures of the same.

12. The method of 10, wherein oxygen is removed by venting the solutionwith hydrogen.

We claim:
 1. A method of producing 2,4-dihydroxybutyrate (DHB),comprising: mixing a four carbon sugar and a hydroxide salt in solution,wherein the four carbon sugar is erythrulose.
 2. The method of claim 1,wherein the temperature of the solution is maintained below 100° C. 3.The method of claim 1, wherein the four carbon sugar is dilutedsufficiently to result in a molar yield of DHB that is greater than 40%.4. The method of claim 1, wherein the four carbon sugar is diluted witha solution containing DHB.
 5. The method of claim 1, wherein the methodis performed in a continuous reactor system.
 6. The method of claim 1,wherein the hydroxide concentration of the solution is between 0.1 M and4 M.
 7. The method of claim 6, wherein the temperature of the solutionis maintained below 100° C.
 8. The method of claim 7, wherein the fourcarbon sugar is diluted with a solution containing DHB.
 9. The method ofclaim 1, further comprising removing oxygen from the solution.
 10. Themethod of claim 9, wherein oxygen is removed by venting the solutionwith a gas selected from the group consisting of: nitrogen, argon, andmixtures thereof.
 11. The method of claim 9, wherein oxygen is removedby venting the solution with hydrogen.
 12. The method of claim 11,wherein the temperature of the solution is maintained below 100° C. 13.The method of claim 12, wherein the erythrulose is diluted with asolution containing one or more other organic acid salts.
 14. The methodof claim 13, wherein the method is performed in a continuous reactorsystem.
 15. A method of producing 2,4-dihydroxybutyrate, comprising:mixing a four carbon sugar and a hydroxide salt in solution; andremoving oxygen from the solution by venting the solution with hydrogen,wherein the four carbon sugar is erythrulose.
 16. The method of claim 1,wherein the four carbon sugar is D-(−)-erythrulose, L(+)-erythrulose,D-erythrulose, L-erythrulose, or any combination thereof.
 17. The methodof claim 1, wherein the four carbon sugar is D-(−)-erythrulose,L(+)-erythrulose, D-erythrulose, L-erythrulose, a salt thereof, ahydrate thereof, or any combination thereof.
 18. The method of claim 15,wherein the four carbon sugar is D-(−)-erythrulose, L(+)-erythrulose,D-erythrulose, L-erythrulose, or any combination thereof.
 19. The methodof claim 15, wherein the four carbon sugar is D-(−)-erythrulose,L(+)-erythrulose, D-erythrulose, L-erythrulose, a salt thereof, ahydrate thereof, or any combination thereof.
 20. The method of claim 15,wherein the temperature of the solution is maintained below 100° C.